Effects of apamin and rapamycin on S6rp phosphorylation in CINs are reproduced <i>in vivo</i>.

<p>(A) Low magnification image of a rat brain section showing the targeted striatal region bilaterally (red cross). Rats received either synaptic blocker solution (blockers; picrotoxin, 150 µM; CNQX disodium, 1 mM; DL-AP5, 1 mM) or the same solution plus experimental drug(s) (apamin, 100 nM and/or rapamycin, 1 µM) in contralateral hemispheres. (B) Confocal images from a transcardially-fixed rat brain previously injected with blockers (left-side dorsal striatum, Left DMS) and blockers plus apamin (right-side dorsal striatum, Right DMS) showing double staining for ChAT and p-Ser^240–244-S6rp. Top panels are low-magnification images showing several cholinergic interneurons in the same focal plane. Bottom panels are higher magnification images showing p-S6rp signal intensity in CINs from the left (blockers) and right (blockers + apamin) striata of the same animal. Insets show corresponding ChAT staining. (C) Quantification of p-S6rp signal in striatal ChAT immunoreactive neurons after each combination of injections (intensity for each neuron was calculated as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053195#pone-0053195-g002" target="_blank">Figure 2E</a>). A 16 pseudo-color palette LUT highlights the intensity of p-S6rp fluorescence. In the scatterplot, each dot corresponds to one neuron; each color corresponds to a different animal; dashed lines indicate the mean. Fluorescence values are normalized into arbitrary units (a.u.). Data were analyzed with one-way ANOVA (Effect of drug: F_(5,188) = 2.932, p = 0.0142, Bonferroni post hoc: Blockers vs. Blockers+Apamin; p<0.01; 30–35 quantified neurons per condition in 9 rats).